Th17/Treg homeostasis, but not Th1/Th2 homeostasis, is implicated in exacerbation of human bronchial asthma

Xiao-Ling Zou, Zhuang-Gui Chen, Tian-Tuo Zhang, Ding-Yun Feng, Hong-Tao Li, Hai-Ling Yang, Xiao-Ling Zou, Zhuang-Gui Chen, Tian-Tuo Zhang, Ding-Yun Feng, Hong-Tao Li, Hai-Ling Yang

Abstract

Background: Th17 and regulatory T cell (Treg) play crucial roles in the pathogenesis of asthma. However, the association between Th17/Treg homeostasis and asthma exacerbation remains unclear.

Patients and methods: To investigate the role of Th17/Treg bias in asthma exacerbation, 49 asthma patients were enrolled in the current study, of whom 31 had acute asthma exacerbation (exacerbation group) and 18 did not (non-exacerbation group). Meanwhile, 17 healthy subjects were recruited as normal controls (control group). By measuring interleukin (IL)-4, IL-13, interferon (IFN)-γ, IL-10, and IL-17A levels in plasma using enzyme-linked immunosorbent assay (ELISA) and determining the mRNA expression of T-bet, GATA-3, forkhead/winged helix transcription factor (Foxp3), and receptor-related orphan receptor γt (RORγt) in peripheral blood mononuclear cells (PBMCs) by quantitative real-time PCR.

Results: We found that IL-17A/IL-10 and RORγt/Foxp3 ratios were significantly increased in the exacerbation group compared with that in the non-exacerbation group, while IL-4/IFN-γ and GATA-3/T-bet ratios remained unchanged. Moreover, IL-17A/IL-10 and RORγt/Foxp3 ratios, but not IL-4/IFN-γ or GATA-3/T-bet ratios, negatively correlated with forced expiratory volume in the first second (FEV1)/FEV1pred and Asthma Control Test Questionnaire (ACT) scores in both exacerbation group and non-exacerbation group. In contrast, the IL-4/IFN-γ ratio was negatively correlated with FEV1/FEV1pred and ACT scores only in the non-exacerbation group but not in the exacerbation group, while the ratio of GATA-3/T-bet was correlated with neither FEV1/FEV1pred nor ACT scores in both groups with asthma.

Conclusion: Our results suggest that the homeostasis of the Treg and Th17 cells is broken in asthma exacerbation and correlates with asthma severity and disease control status. The outcome has significant implication in understanding the progression of asthma and providing helpful information for physicians in the diagnosis and treatment of asthma patients.

Keywords: T cell; Th17/Treg; bronchial asthma; exacerbation.

Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
Lung function and ACT scores. Notes: (A) FEV1/FEV1pred levels in three groups. (B) The ACT scores in patients with asthma (*P < 0.05, **P < 0.01). Abbreviations: ACT, Asthma Control Test Questionnaire; FEV1, forced expiratory volume in the first second.
Figure 2
Figure 2
The cytokine levels in plasma. Notes: (A) Cytokine IFN-γ, IL-4, IL-17A, IL-10, and IL-13 expression levels in the three groups. (B) The ratio of IL-4/IFN-γ and IL-17A/IL-10 in plasma. (*P < 0.05 versus control group, #P < 0.05 versus non-exacerbation group, one-way analysis of variance). Abbreviations: IFN-γ, interferon-γ; IL, interleukin.
Figure 3
Figure 3
The transcription factor mRNA levels in plasma. (The mRNA level calculated here is relative to house-keeping gene expression.) Notes: (A) The transcription factor levels in the three groups. (B) The ratio of GATA-3/T-bet and RORγt/Foxp3 in plasma. (*P < 0.05 versus control group, #P < 0.05 versus non-exacerbation group, Mann–Whitney test). Abbreviations: Foxp3, forkhead/winged helix transcription factor 3; GATA-3, GATA binding protein 3; RORγt, retinoic acid–related orphan receptor gamma T; T-bet, T-box expressed in T cells.
Figure 4
Figure 4
Spearman’s correlation analysis between Th17/Treg and Th2/Th1 homeostasis in asthmatic patients. Notes: Correlation analysis between IL-17A/IL-10 and IL-4/IFN-γ (A, r = −0.201, P = 0.278) and RORγt/Foxp3 and GATA-3/T-bet in exacerbation group (B, r = −0.250, P = 0.173) and between IL-17A/IL-10 and IL-4/IFN-γ (C, r = 0.876, P = 0.054) and RORγt/Foxp3 and GATA-3/T-bet (D, r = −0.231, P = 0.338) in non-exacerbation group. P-value less than 0.05 means significant correlation. Abbreviations: Foxp3, forkhead/winged helix transcription factor 3; GATA-3, GATA binding protein 3; IFN-γ, interferon-γ; IL, interleukin; RORγt, retinoic acid–related orphan receptor gamma T; T-bet, T-box expressed in T cells.
Figure 5
Figure 5
Spearman’s correlation analysis between cytokine expression ratios and disease severity in exacerbation group. Notes: Correlation analysis between IL-17A/IL-10 and FEV1/FEV1pred (A, r = −0.769, P = 0.001) and ACT scores (B, r = −0.487, P = 0.016) and between IL-4/IFN-γ and FEV1/FEV1pred (C, r = 0.307, P = 0.093) and ACT scores (D, r = 0.415, P = 0.075). P-value less than 0.05 means significant correlation. Abbreviations: ACT, Asthma Control Test Questionnaire; FEV1, forced expiratory volume in the first second; IFN-γ, interferon-γ; IL, interleukin.
Figure 6
Figure 6
Spearman’s correlation analysis between cytokine expression ratios and disease severity in non-exacerbation group. Notes: Correlation analysis between IL-17A/IL-10 and FEV1/FEV1pred (A, r = −0.965, P = 0.003) and ACT scores (B, r = −0.667, P = 0.029) and between IL-4/IFN-γ and FEV1/FEV1pred (C, r = −0.842, P = 0.004) and ACT scores (D, r = −0.467, P = 0.044). P-value less than 0.05 means significant correlation. Abbreviations: ACT, Asthma Control Test Questionnaire; FEV1, forced expiratory volume in the first second; IFN-γ, interferon-γ; IL, interleukin.
Figure 7
Figure 7
Spearman’s correlation analysis between transcription factor mRNA levels and disease severity in exacerbation group. Notes: Correlation analysis between RORγt/Foxp3 and FEV1/FEV1pred (A, r = −0.725, P = 0.016) and ACT scores (B, r = −0.632, P = 0.015) and between GATA-3/T-bet and FEV1/FEV1pred (C, r = 0.172, P = 0.355) and ACT scores (D, r = 0.183, P = 0.324). P-value less than 0.05 means significant correlation. Abbreviations: ACT, Asthma Control Test Questionnaire; FEV1, forced expiratory volume in the first second; Foxp3, forkhead/winged helix transcription factor; GATA-3, GATA binding protein 3; RORγt, retinoic acid–related orphan receptor gamma T; T-bet, T-box expressed in T cells.
Figure 8
Figure 8
Spearman’s correlation analysis between transcription factor mRNA levels and disease severity in non-exacerbation group. Notes: Correlation analysis between RORγt/Foxp3 and FEV1/FEV1pred (A, r = −0.758, P = 0.028) and ACT scores (B, r = −0.415, P = 0.086) and between GATA-3/T-bet and FEV1/FEV1pred (C, r = 0.292, P = 0.239) and ACT scores (D, r = 0.301, P = 0.225). P-value less than 0.05 means significant correlation. Abbreviations: ACT, Asthma Control Test Questionnaire; FEV1, forced expiratory volume in the first second; Foxp3, forkhead/winged helix transcription factor; GATA-3, GATA binding protein 3; RORγt, retinoic acid–related orphan receptor gamma T; T-bet, T-box expressed in T cells.

References

    1. Iwata A, Kawashima S, Kobayashi M, et al. Th2-type inflammation instructs inflammatory dendritic cells to induce airway hyperreactivity. Int Immunol. 2014;26(2):103–114.
    1. Hirose K, Iwata A, Tamachi T, Nakajima H. Allergic airway inflammation: key players beyond the Th2 cell pathway. Immunol Rev. 2017;278(1):145–161.
    1. Yagi R, Zhu J, Paul WE. An updated view on transcription factor GATA3-mediated regulation of Th1 and Th2 cell differentiation. Int Immunol. 2011;23(7):415–420.
    1. Bosnjak B, Stelzmueller B, Erb KJ, Epstein MM, Klaus J, Michelle M. Treatment of allergic asthma: modulation of Th2 cells and their responses. Respir Res. 2011;12:114.
    1. Afshar R, Medoff BD, Luster AD. Allergic asthma: a tale of many T cells. Clin Exp Allergy. 2008;38(12):1847–1857.
    1. Talaat RM, Mohamed SF, Bassyouni IH, Raouf AA. Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity. Cytokine. 2015;72(2):146–153.
    1. Zhang H, Kong H, Zeng X, Guo L, Sun X, He S. Subsets of regulatory T cells and their roles in allergy. J Transl Med. 2014;12:125.
    1. Guzmán-Flores JM, Portales-Pérez DP. Mechanisms of suppression of regulatory T-cells (Treg) Gac Med Mex. 2013;149(6):630–638.
    1. Nakada EM, Shan J, Kinyanjui MW, Fixman ED. Adjuvant-dependent regulation of interleukin-17 expressing γδ T cells and inhibition of Th2 responses in allergic airways disease. Respir Res. 2014;15(1):90.
    1. Zhu J, Liu X, Wang W, Ouyang X, Zheng W, Wang Q. Altered expression of regulatory T and Th17 cells in murine bronchial asthma. Exp Ther Med. 2017;14(1):714–722.
    1. Jiang H, Wu X, Zhu H, Xie Y, Tang S, Jiang Y. FOXP3(+)Treg/Th17 cell imbalance in lung tissues of mice with asthma. Int J Clin Exp Med. 2015;8(3):4158–4163.
    1. Huang X, Chen Y, Zhang F, Yang Q, Zhang G. Peripheral Th17/Treg cell-mediated immunity imbalance in allergic rhinitis patients. Braz J Otorhinolaryngo. 2014;80(2):152–155.
    1. Shi YH, Shi GC, Wan HY, et al. Coexistence of Th1/Th2 and Th17/Treg imbalances in patients with allergic asthma. Chinese Medical Journal. 2011;124(13):1951–1956.
    1. Wei B, Zhag H, Li L, Li M, Shang Y. T Helper 17 Cells and Regulatory T-Cell Imbalance in Paediatric Patients with Asthma. J Int Med Res. 2011;39(4):1293–1305.
    1. Hartl D, Koller B, Mehlhorn AT, et al. Quantitative and functional impairment of pulmonary CD4+CD25hi regulatory T cells in pediatric asthma. J Allergy Clin Immunol. 2007;119(5):1258–1266.
    1. Strickland DH, Stumbles PA, Zosky GR, et al. Reversal of airway hyperresponsiveness by induction of airway mucosal CD4+ CD25+ regulatory T cells. J Exp Med. 2006;203(12):2649–2660.
    1. Park BS, Hong GU, Ro JY, Jy R. Foxp3+-Treg Cells Enhanced by Repeated Low-Dose Gamma-Irradiation Attenuate Ovalbumin-Induced Allergic Asthma in Mice. Radiat Res. 2013;179(5):570–583583.
    1. Wilson RH, Whitehead GS, Nakano H, Free ME, Kolls JK, Cook DN. Allergic Sensitization through the Airway Primes Th17-dependent Neutrophilia and Airway Hyperresponsiveness. Am J Respir Crit Care Med. 2009;180(8):720–730.
    1. Tao B, Ruan G, Wang D, Li Y, Wang Z. Imbalance of Peripheral Th17 and Regulatory T Cells in Children with Allergic Rhinitis and Bronchial Asthma. Iran J Allergy Asthma Immunol. 2015;14(3):273–279.
    1. Fuss IJ, Kanof ME, Smith PD, Zola H. Isolation of whole mononuclear cells from peripheral blood and cord blood. Curr Protoc Immunol. 2009 Chapter 7–Unit7.1.
    1. Zhang W, Zhang X, Sheng A, et al. γ-Secretase Inhibitor Alleviates Acute Airway Inflammation of Allergic Asthma in Mice by Down-regulating Th17 Cell Differentiation. Mediators Inflamm. 2015;2015(1):258168, 7.
    1. Zhao J, Lloyd CM, Noble A. Th17 responses in chronic allergic airway inflammation abrogate regulatory T-cell-mediated tolerance and contribute to airway remodeling. Mucosal Immunol. 2013;6(2):335–346.
    1. Zemedkun K, Woldemichael K, Tefera G. Assessing Control of Asthma in Jush, Jimma, South West Ethiopia. Ethiop J Health Sci. 2014;24(1):49–58.
    1. Manoharan A, Anderson WJ, Lipworth J, Lipworth BJ. Assessment of Spirometry and Impulse Oscillometry in Relation to Asthma Control. Lung. 2015;193(1):47–51.
    1. Chae EJ, Kim T-B, Cho YS, et al. Airway Measurement for Airway Remodeling Defined by Post-Bronchodilator FEV1/FVC in Asthma: Investigation Using Inspiration-Expiration Computed Tomography. Allergy, Asthma and Immunology Research. 2011;3(2):111–117.
    1. Zhao W, Yun X, Zhi W. Neutralization of interleukin-17 suppresses allergic rhinitis symptoms by downregulating Th2 and Th17 responses and upregulating the Treg response. Oncotarget. 2017;8(14):22361–22369.
    1. Wang L, Wan H, Tang W, et al. Critical roles of adenosine A2A receptor in regulating the balance of Treg/Th17 cells in allergic asthma. Clin Respir J. 2018;12(1):149–157.

Source: PubMed

Спонсоры и соавторы

Заболевания

Медикаментозные вмешательства

Подписаться